钱秀娟,
张世鹏,
谢攀,
刘长仲^,
甘肃农业大学植物保护学院/甘肃省农作物病虫害生物防治工程实验室 兰州 730070
基金项目: 甘肃农业大学科技创新基金-学科建设专项GAU-XKJS-151
甘肃省科技计划项目18JR3RA162

详细信息

作者简介:钱秀娟, 主要研究方向为害虫生物防治。E-mail:qianxj@gsau.edu.cn

通讯作者:刘长仲, 主要研究方向为昆虫生态及害虫治理。E-mail:liuchzh@gsau.edu.cn

中图分类号:S476+15

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出版历程

收稿日期:2018-06-16
录用日期:2018-10-09
刊出日期:2019-06-01

Effects of UV-B radiation on entomopathogenic nematode survival and pathogenicity

QIAN Xiujuan,
ZHANG Shipeng,
XIE Pan,
LIU Changzhong^,
College of Plant Protection of Gansu Agriculture University/Biocontrol Engineering Laboratory of Crop Diseases and Pests of Gansu Province, Lanzhou 730070, China
Funds: the Science and Technology Innovation Fund of Gansu Agricultural UniversityGAU-XKJS-151
The Science and Technology Fund of Gansu Province18JR3RA162

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Corresponding author:LIU Changzhong, E-mail:liuchzh@gsau.edu.cn

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摘要
摘要:为探明紫外辐射对昆虫病原线虫（entomopathogenic nematodes，EPN）的作用，以甘肃省分离得到的昆虫病原线虫为研究材料，用室内模拟生测法，测定了经20 W、40 W紫外线辐射后，斯氏属（Steinernema feltiae，S.kraussei）和异小杆属（Heterorhabditis brevicaudis，H.megidis）4种昆虫病原线虫9个品系的存活率，并以大蜡螟（Galleria mellonella）为供试昆虫，采用改良的ONE ON ONE法，测定了其对大蜡螟的致病力。结果表明：经UV-B辐射后供试的9个昆虫病原线虫品系存活率未发生显著变化（P>0.05）。20 W紫外辐射30 min后，S.feltiae 0639C和0663TD 2品系对大蜡螟的致病力提高，其中S.felitiae 0639C致病力显著升高（P < 0.05），侵染32 h后，寄主昆虫死亡率由13.3%升高到73.3%；S.felitiae 0661YM品系的致病力显著降低（P < 0.05），侵染32 h后，寄主昆虫死亡率率由66.7%降低到0；其余各品系致病力均无显著变化。不同辐射时长和辐射强度对S.felitiae 0639C和0663TD引起寄主昆虫死亡的时间有影响，较对照组而言，经辐射预处理后，2品系线虫引起寄主昆虫死亡的时间均提前，其中S.felitiae 0639C随着辐射强度的增大引起寄主昆虫开始死亡时间提前，而S.felitiae 0663TD随着辐射强度增加引起寄主昆虫开始死亡时间延后。不同紫外辐射时长对S.felitiae 0663TD和S.felitiae 0639C致病力无显著影响，30 min、60 min、90 min、120 min辐射处理之间差异不显著（P>0.05）。研究表明，9个供试昆虫病原线虫品系均对UV-B辐射具有一定的耐受能力；20 W UV-B辐射30 min，能刺激S.felitiae 0663TD和S.felitiae 0639C侵染和致死寄主昆虫；一定的辐射时长内（120 min），辐射时长对S.felitiae 0663TD和S.felitiae 0639C的致病力无影响。
关键词:昆虫病原线虫/
UV-B辐射/
大蜡螟/
存活率/
致病力/
非生物胁迫
Abstract:Entomopathogenic nematodes (EPN) possess many positive attributes as biocontrol agents. They are safe to humans and generally safe to other non-target organisms. To investigate the effects of ultraviolet radiation exposure on EPN, we studied the effects of 20 or 40 W UV-B radiation for different time lengths (30, 60, 90, or 120 min) on the virulence of EPN infective juveniles (IJs) to Galleria mellonella, including 9 strains from 4 species (Steinernema feltiae, S. kraussei, Heterorhabditis brevicaudis, and H. megidis) isolated from Gansu Province. The results showed that there were no differences among the strains in the survival rate of IJs with and without 30 min UV-B irradiation. The virulence of IJs varied among strains after exposure to 20 W ultraviolet radiation for 30 min: virulence of two strains (S. felitiae 0639C and S. feltiae 0663TD) was increased (P < 0.05), that of one strain (S. feltiae 0661YM) decreased (P < 0.05), and that of all others (H. brevicaudis 0641TY, H. megidis 0627M, S. felitiae 0639C, S. felitiae 0619HT, S. kraussei 0663HT, and S. kraussei 0657L) showed no significantly change (P > 0.05). For example, when measured at 32 h after infection, the mortality rate of host insects infected by S. felitiae 0639C increased significantly, from 13.3% to 73.3% (P < 0.05), whereas that of insects infected by S. felitiae 0661YM decreased significantly from 66.7% to zero (P < 0.05). At the same time, treating S. felitiae 0639C and 0663TD with UV affected host mortality 24 h after exposure to IJs:for S. felitiae 0639C, host mortality at 24 h was greater in the 40 W treatment than in either control or the 20 W treatment; for S. felitiae 0663TD, host mortality at 24 h was greater in the 20 W treatment than in either the control or the 40 W treatment. Ultraviolet radiation duration (30 min, 60 min, 90 min, or 120 min exposure) had no effect on the pathogenicity of S. felitiae 0663TD or S. felitiae 0639C (P > 0.05). Therefore, the following conclusions were drawn:All of the 9 tested strains of EPN isolated from Gansu Province had some tolerance to UV-B radiation, and, in particular, S. felitiae 0622L had a high tolerance to ultraviolet radiation. In addition, ultraviolet radiation had a variable impact on the pathogenicity of EPN in Gansu Province, as there were differences among different species and strains. For example, 20 W UV-B radiation for 30 min enhanced the pathogenicity of S. felitiae 0663TD, whereas it significantly weakened that of S. felitiae 0661YM. Ultraviolet radiation stimulated S. felitiae 0663TD and S. felitiae 0639C to infect and kill host insects more quickly, although the radiation intensity that optimized the 24-h mortality rate varied between the two strains. After 48 h, however, there was no significant difference in the pathogenicity of S. felitiae 0663TD and S. felitiae 0639C between the different radiation intensities (20 and 40 W). Additionally, IJs of S. felitiae 0663TD and S. felitiae 0639C both had high pathogenicity after 48-72 h, regardless of the radiation intensity (20 W or 40 W) or the duration of exposure (30, 60, 90, or 120 min). These findings will shed more light on using entomopathogenic nematodes under ultraviolet radiation stress.
Key words:Entomopathogenic nematodes/
Ultraviolet radiation/
Galleria mellonella/
Survival rate/
Pathogenicity/
Abiotic stress

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图1UV-B辐射强度与处理时间对处理后不同时间S. felitiae 0639C(a)和0663TD(b)致病力的影响
Figure1.Effects of UV-B radiation time and intensity on pathogenicity of S. felitiae 0639C (a) and 0663TD (b) after different times of treatment


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表1不同时间不同强度UV-B辐射对不同昆虫病原线虫品系存活率的影响
Table1.Entomopathogenic nematodes infective juveniles (IJs) survival rate at different ultraviolet radiation intensities under different exposure time length

线虫种 Species	品系 Strain	处理 Treatment	存活率 Survival rate (%)
			0.5 h	1 h	1.5 h	2 h	3 h	4 h	5 h	6 h
Steinernema feltiae	0639C	CK	95.3±2.3aA	95.3±2.3aA	95.3±2.3aA	95.3±2.3aA	95.3±2.3aA	95.3±2.3aA	95.1±2.3aA	95.3±2.3aA
		20W	92.3±1.7aA	90.0±0.5aA	87.1±2.1aA	83.3±2.3aA	56.1±2.1bABC	30.1±2.8bBC	12.6±0.5bB	0bB
		40W	91.7±2.1aA	77.7±0.4bAB	63.1±0.4bA	60.3±1.1bAB	50.5±2.8bABC	26.7±2.4bBC	3.4±0.5bB	0bB
	0661YM	CK	91.6±3.4aA	91.6±3.4aA	91.6±3.4aA	91.6±3.4aA	91.6±3.4aA	91.6±3.4aA	91.6±3.5aA	91.6±3.4aA
		20W	89.9±4.1aA	86.4±1.5aA	84.6±2.1aA	84.6±3.6aA	34.5±2.6bBC	10.2±0.4bC	3.9±0.2bB	0bB
		40W	89.1±3.7aA	81.1±0.9aA	75.1±3.5bAB	79.3±4.6bA	30.1±5.3bBC	4.9±1.0bC	0bB	0bB
	0619HT	CK	98.7±0.5aA	98.8±0.5aA	98.8±0.5aA	98.8±0.5aA	98.8±0.5aA	98.8±0.5aA	98.7±0.5aA	98.8±0.5aA
		20W	96.4±0.7aA	94.3±3.1aA	94.1±2.7aA	92.6±3.1aA	42.2±1.8bBC	15.2±5.7bBC	5.2±0.8bB	2.7±1.3bB
		40W	95.3±2.5aA	89.7±3.5aA	80.6±4.8bA	78.4±2.1bAB	31.3±3.5bBC	8.7±2.5bC	2.1±0.5bB	1.7±0.4bB
	0622L	CK	83.6±3.4aA	83.6±3.4aA	83.6±3.4aA	83.6±3.4aA	83.6±3.4aA	83.6±3.4aA	83.5±3.5aA	83.6±3.4aA
		20W	80.3±4.5aA	69.0±3.7aAB	67.6±2.5aAB	59.3±1.3bAB	39.3±2.3bBC	12.5±3.6bC	3.6±1.2bB	3.2±0.6bB
		40W	69.9±3.6bA	50.1±4.6bB	48.4±3.3bB	40.4±4.2bB	34.5±1.4bBC	9.3±3.2bC	3.4±0.8bB	1.4±0.3bB
	0663TD	CK	93.3±2.4aA	93.3±2.4aA	93.3±2.4aA	93.3±2.4aA	93.3±2.4aA	93.3±2.4aA	93.3±2.4aA	93.3±2.4aA
		20W	91.8±2.6aA	81.2±3.5abA	78.9±4.1abAB	77.8±2.5abAB	45.3±3.6bBC	24.6±3.1bBC	10.2±2.4bB	1.2±0.7bB
		40W	91.0±1.3aA	68.7±2.8bAB	68.1±4.6bAB	64.8±3.4bAB	41.8±2.5bBC	20.3±4.1bBC	8.8±1.4bB	0.9±0.5bB
Steinernema kraussei	0657L	CK	99.3±0.2aA	99.3±0.2aA	99.3±0.2aA	99.3±0.2aA	99.3±0.2aA	99.3±0.2aA	99.2±0.2aA	99.3±0.2aA
		20W	98.7±0.9aA	97.4±0.9aA	94.8±4.1aA	96.3±2.1aA	83.3±9.9aA	23.6±6.3bBC	5.5±1.2bB	3.4±4.2bB
		40W	96.2±1.4aA	92.4±2.3aA	91.5±2.3aA	90.8±1.7aA	74.2±4.1aAB	17.4±2.3bC	4.1±0.7bB	1.7±0.9bB
	0663HT	CK	83.3±6.9aA	83.3±6.9aA	83.3±6.9aA	83.3±6.9aA	83.3±6.9aA	83.3±6.9aA	83.3±6.8aA	83.3±6.9aA
		20W	82.1±5.7aA	76.3±1.3aAB	67.7±0.9abAB	50.4±0.6bB	31.4±1.3bBC	9.6±0.4bC	1.6±0.3bB	0bB
		40W	81.8±1.3aA	71.5±2.4bAB	60.1±2.4bAB	44.1±1.4bB	22.4±0.5bC	3.6±1.0bC	0bB	0bB
Heterorhabditis brevicaudis	0627M	CK	87.7±5.8aA	87.7±5.8aA	87.7±5.8aA	87.7±5.8aA	87.7±5.8aA	87.7±5.8aA	87.7±5.8aA	87.7±5.8aA
		20W	86.4±2.8aA	84.3±1.3aA	80.6±4.1aA	74.2±2.4abAB	38.4±0.5bBC	20.6±2.3bBC	6.5±4.1bB	1.7±0.7bB
		40W	86.1±3.1aA	83.3±3.4aA	78.9±0.9aAB	70.6±4.1bAB	30.6±3.4bBC	12.3±1.2bC	4.1±0.7bB	0bB
大异小杆线虫 Heterorhabditis megidis	0641TY	CK	98.8±1.1aA	98.8±1.1aA	98.8±1.1aA	98.8±1.1aA	98.8±1.1aA	98.8±1.1aA	98.8±1.1aA	98.8±1.1aA
		20W	98.2±0.5aA	98.1±0.3aA	97.9±0.3aA	97.3±0.1aA	85.4±3.3abA	53.6±5.7bB	10.7±3.9bB	5.0±2.6bB
		40W	97.1±1.0aA	84.2±0.7bA	84.2±1.2bA	82.3±1.4bA	79.4±2.7bAB	49.2±3.4bB	6.7±2.6bB	3.7±1.7bB
采用t检验, 不同小写字母表示同一品系在不同处理之间差异显著(P < 0.05), 不同大写字母表示不同品系不同处理之间差异显著(P < 0.05)。Different lowercase letters indicate significant differences among treatments within the same strain, while different capital letters stand for the significant differences among different strains (P < 0.05) by t test.

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表220W UV-B辐射30 min下不同昆虫病原线虫品系对大蜡螟致病力的影响
Table2.Effects of 20 W ultraviolet radiation after 30 min exposure on entomopathogenic nematodes' pathogenicity to Galleria mellonella

线虫种Species	品系Strain	处理Treatment	累积校正死亡率Corrected mortality (%)
			32 h	40 h	48 h	56 h	64 h	72 h	80 h	88 h	96 h	104 h	112 h	120 h
Steinernema feltiae	0639C	CK	13.3± 1.6bCD	20.0± 2.3bBCD	53.3±2.6aABC	73.3± 4.1aABC	100aA	100aA	100aA	100aA	100aA	100aA	100aA	100aA
		UV	73.3± 4.1aA	86.6± 3.4aA	86.6±3.4aAB	100.0aA	100aA	100aA	100aA	100aA	100aA	100aA	100aA	100aA
	0661YM	CK	66.7± 3.8aAB	66.6± 3.8aAB	80.0±6.7aABC	100.0aA	100aA	100aA	100aA	100aA	100aA	100aA	100aA	100aA
		UV	0bD	0bD	20.0±2.3bBC	20.0± 2.3bB	26.67bB	26.67bB	46.7± 4.1bA	66.7± 3.8aA	80.0± 6.7aAB	80± 6.7aA	93.3± 1.7aA	100aA
	0619HT	CK	53.3±2.6 aABC	86.6± 3.4aAB	93.3±1.7aA	100.0aA	100aA	100aA	100aA	100aA	100aA	100aA	100aA	100aA
		UV	46.7±4.1 aABCD	46.7± 4.1aABC	60.0±3.1aABC	60.0± 3.1aAB	73.3± 4.1aAB	73.3± 4.1aAB	73.3± 4.1aA	80± 6.7aA	93.3± 1.7aA	93.3± 1.7aA	93.3± 1.7aA	100aA
	0622L	CK	13.3±1.6 aCD	26.7± 3.9aABC	66.7±3.8aABC	73.3± 4.1aAB	73.3± 4.1aAB	80.0± 6.7aAB	80.0± 6.7aA	80.0± 6.7aA	80.0± 6.7aA	86.7± 3.4aA	100aA	100aA
		UV	26.7±3.9 aABCD	46.7±4.1aABC	53.3±2.6aABC	73.3± 4.1aAB	86.7± 3.4aAB	86.7± 3.4aAB	86.7± 3.4aA	86.7± 3.4aA	86.7± 3.4aA	100aA	100aA	100aA
	0663TD	CK	66.6±3.8 aAB	66.7± 3.8aABC	66.7±3.8aABC	93.3± 1.7Aa	93.3± 3.4aA	93.3± 1.7aA	100aA	100aA	100aA	100aA	100aA	100aA
		UV	93.3± 1.7aA	100.0aA	100.0aA	100aA	100aA	100aA	100aA	100aA	100aA	100aA	100aA	100aA
Steinernema kraussei	0657L	CK	0aD	46.7± 4.1aABC	73.3±4.1aABC	73.3± 4.1aAB	80.0± 6.7aAB	93.3± 1.7aA	93.3± 1.7aA	100aA	100aA	100aA	100aA	100aA
		UV	0aD	33.3± 1.7aABC	33.3±1.7aABC	46.6± 4.1aAB	66.7± 3.8aAB	73.3± 4.1aAB	73.3± 4.1aA	86.7± 3.4aA	100aA	100aA	100aA	100aA
	0663HT	CK	40.0±1.9 aABCD	60.0± 2.1aABC	60.0±2.1aABC	60.0± 2.1aAB	66.7± 3.8aAB	73.3± 4.1aAB	86.7± 3.4aA	100aA	100aA	100aA	100aA	100aA
		UV	20.0±2.3 aBCD	53.3± 2.6aABC	66.7±3.8aABC	66.7± 3.8aAB	66.7± 3.8aAB	66.7± 3.8aAB	80.0± 6.7aA	86.7± 3.4aA	93.3± 1.7aA	100aA	100aA	100aA
Heterorhabditis brevicaudis	0627M	CK	6.7± 1.1aCD	20.0± 2.3aBC	33.3±2.9aABC	60.0± 3.1aAB	80.0± 6.7aAB	80.0± 6.7aA	100aA	100aA	100aA	100aA	100aA	100aA
		UV	6.7± 1.1aCD	40.0± 1.9aABC	53.3±2.6aABC	60.0± 3.1aAB	66.7± 3.8aAB	73.3± 4.1aA	86.7± 3.4aA	100aA	100aA	100aA	100aA	100aA
大异小杆线虫Heterorhabditis megidis	0641TY	CK	26.7±3.9 aABCD	60.0± 3.1aABC	80.0±6.7aABC	80.0± 6.7aAB	86.7± 3.4aAB	93.3± 1.7aA	93.3± 1.7aA	93.3± 1.7aA	93.3± 1.7aA	100aA	100aA	100aA
		UV	6.7±1.1 aCD	53.3±2.6aABC	73.3±4.1aABC	80.0± 6.7aAB	86.7± 3.4aAB	86.7± 3.4aA	86.7± 3.4aA	86.7± 3.4aA	93.3± 1.7aA	93.3± 1.7aA	93.3± 1.7aA	100aA
采用t检验, 不同小写字母表示同一品系在不同处理之间差异显著(P < 0.05), 不同大写字母表示不同品系不同处理之间差异显著(P < 0.05)。Different lowercase letters indicate significant differences among treatments within the same strain, while different capital letters stand for the significant differences among different strains (P < 0.05) by t test.

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